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反相液相色谱在蛋白质及多肽分离分析中的应用 总被引:22,自引:0,他引:22
反相液相色谱是一种以疏水作用为基础的色谱分离模式。由于它具有分辨率高、重复性好、回收率高等优点,在蛋白质及多肽的分离分析中得到了极为广泛的应用。本文简要介绍了反相液相色谱及其分离机理,对其在蛋白质和多肽研究中的应用如分离纯化、肽图分析、酶活测定、构象变化检测及疏水作用研究等作系统综述,并展望了反相液相色谱在这一领域的发展前景。 相似文献
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唑来膦酸及其有关化合物的反相离子对高效液相色谱分离 总被引:6,自引:0,他引:6
采用反相离子对高效液相色谱法研究了唑来膦酸及其有关化合物的色谱分析与分离方法。优化的分离条件:以Hypersil C8柱为固定相,以甲醇-5 mmol/L磷酸二氢钠缓冲液(含6 mmol/L四丁基溴化铵溶液,用氢氧化钠溶液调节pH至7.0)(体积比为20∶80)为流动相,等度洗脱,流速为1.0 mL/min,紫外检测波长为220 nm,柱温为室温。在该色谱条件下,唑来膦酸与有关化合物(包括其合成过程中残余的原料咪唑乙酸和其他氧化分解产物)的分离良好,与保留时间最接近的杂质峰的分离度大于2.5。该方法不需进 相似文献
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《液相色谱法及相关技术杂志》2012,35(7):919-933
Abstract This paper describes the use of high performance liquid chromatography (HPLC) for the rapid analysis and purification of insulin-related peptides prepared by solid-phase synthetic procedures. Examples include the bovine insulin C-peptide (34–45), the porcine insulin C-peptide (41–53) and the insulin B-chain fragment (22–27). Chromatographic elution systems containing reducing reagents like B-mercaptoethanol allow the direct analysis of insulin reduction products. Similar systems should allow the rapid analysis of disulphide bond pairing patterns in appropriate polypeptides and proteins either directly or following proteolytic digestion. Reversed-phase high performance liquid chromatography (RP-HPLC) is a versatile and rapid technique useful for the analysis and purification of biological substances. In a series of recent publication1–4 we have described methods for the analysis of underivatised amino acids, peptides and proteins on reversed-phase packings using ion-pairing or stationary phase modifying reagents as components of the mobile phase. These studies demonstrated that excellent resolution of closely related peptides can be achieved under a variety of elution conditions. The addition of low levels of phosphoric acid, inorganic or organic phosphates to a mobile phase (generally water-organic solvent mixtures), in particular, allows rapid and reproducible analysis of peptidic compounds with high sensitivity detection at wavelengths down to 190nm5,6. It is the purpose of this report to show that these chromatographic conditions allow the facile analysis, and purification, of a variety of insulin-related peptides. 相似文献
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L. A. Kartsova A. A. Sidorova V. A. Kazakov E. A. Bessonova A. Ya. Yashin 《Journal of Analytical Chemistry》2004,59(8):737-741
A method is proposed for the electrophoretic determination of adrenaline, noradrenaline, and dopamine with mass-spectrometric or UV detection. A procedure is proposed for the sample preparation of biological fluids with the use of solid-phase extraction on alumina. A comparative assessment of the determination of catecholamines by capillary electrophoresis and reversed-phase high-performance liquid chromatography was performed. 相似文献
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《液相色谱法及相关技术杂志》2012,35(6):1229-1247
Abstract In this study, we describe a novel approach to preparative liquid chromatography which takes advantage of the different relative hydrophobicities of components of a sample mixture, so that when a column is optimally loaded with an aqueous solution of the sample mixture, there is competition among the sample components for the adsorption sites on the hydrophobic stationary phase. The more hydrophobic components compete more successfully for these sites than more hydrophilic components, which are displaced and immediately eluted from the column. Thus, the major separation takes place in water. Subsequent treatment with an aqueous organic modifier is only required to wash retained components off the column and takes no part in the major separation process. This approach was applied to the preparative purification of mixtures of closely-related peptides, representing the crude peptide mixtures typically obtained from solid-phase peptide synthesis. The excellent separation profiles and high yields of pure peptide products on analytical columns reported in this study demonstrate that this methodology has great potential for preparative separation of a major component from hydrophilic and/or hydrophobic impurities. 相似文献
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Eight new carbon adsorbents derived from polyimide copolymer of 4,4'-bis (maleimododiphenyl)methane and divinylbenzene (BM-DVB) were prepared using a variety of methods. The influence of carbonization, steam or phosphoric acid activation on chemical structure and texture of the carbons has been studied. Adsorption-desorption properties of the carbons were estimated based on recovery determinations for, phenol, its chlorinated derivatives and naphthalene from aqueous solution. The results showed that the best adsorption-desorption properties have polymer-based carbons obtained by high temperature steam activation.Revised: 13 February 2006 and 3 April 2006 相似文献
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Nikulicheva O. N. Tikhova V. D. Fadeeva V. P. 《Russian Journal of Applied Chemistry》2002,75(4):659-661
Separation of a mixture of naphthalene derivatives by reversed-phase high-performance liquid chromatography with UV detection was studied. Chromatographic characteristics were calculated for nine compounds, including naphthalene. 相似文献
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反相毛细管电色谱分离山酮类化合物 总被引:1,自引:0,他引:1
目前,毛细管电色谱(CEC)在中草药分析中显示出一些独特的优势,如:CEC流动相相对简单、用量少、峰容量大、更易与质谱(MS)联用。特别是植物提取物中活性成分复杂,需要对色谱峰进行精确的指认并控制峰纯度;采用CEC—MS方法可在线提供中药中各种化合物的质谱图。此外,有许多天然产物在中药中的含量极低,MS的高灵敏度可弥补紫外检测器灵敏度较低的缺陷。因此,研究CEC分离中药成分的选择性问题具有重要的实际意义。蝉翼藤(Securida cainappendiculata Hassk.)为远志科远志族蝉翼藤属攀援灌木,其根茎叶有多种药效,其化学成分主要为黄酮、山酮(XAN)和皂甙。从蝉翼藤中已分离出的10个XAN类成分(结构见图1)。具有显著的抗氧化和保护神经细胞作用。我们曾用毛细管区带电泳(CZE)、高效液相色谱(HPLC)、毛细管胶束电动色谱(MEKC)和毛细管微乳电动色谱(MEEKC)对这10个XAN进行分离,并就电泳条件的变化对分离选择性的影响进行系统研究。本文采用CEC法对这10个XAN成分进行分离,并对其色谱分离条件进行优化研究,以期为今后CEC与MS联用分析中药成分打下基础。 相似文献
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《Analytical letters》2012,45(9-10):1037-1052
Abstract A high performance liquid chromatographic method for the rapid separation and quantitative determination of five barbiturates in pharmaceutical preparations and body fluids has been developed. A C18 reversed phase column was used and the barbiturates were detected at 235nm. A number of eluting systems were examined, the most suitable of them being, Ethanol:Propanol:Methanol:Water (26:5:29:40) at pH = 8.8. 相似文献
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G. D. Brykina S. M. Matusova O. A. Shpigun T. V. Zarubina G. A. Zheltukhina R. P. Evstigneeva 《Journal of Analytical Chemistry》2004,59(3):266-269
The chromatographic retention of some hemin-based peptides on the Diasphere-110-C18 sorbent modified with zinc octa-4,5-carboxyphthalocyaninate was studied. It was demonstrated that this stationary phase is more selective for hemin-peptides than Nucleosil-C18 and Silasorb-600. The conditions of the separation of mixtures of hemin-based peptides were determined, and the parameters of their chromatographic separation were calculated. It was shown that the retention parameters are related to the structure of hemin-based peptides. 相似文献
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《液相色谱法及相关技术杂志》2012,35(16):2927-2947
Abstract The retention behaviour of eleven derivatives of counarin was investigated by reversed-phase (RP) thin-layer and high performance liquid chromatography. The linear relationships between log k'or RM values and the content of organic modifier in the aqueous mobile phase obtained for wide composition ranges indicate that the plots can be used to determine RMw and log kw values by extrapolation to pure water. The effects of individual substituants on the retention and the correlation between TLC and HPLC data was analysed. 相似文献